Survey of the occurrence of aflatoxin M1 in dairy products marketed in Italy: second year of observation.

During 1996, 161 samples of milk, 92 samples of dry milk for infant formula and 120 samples of yoghurt, were randomly collected in supermarkets and drug stores in four big Italian cities, and checked for aflatoxin M1 (AFM1) by immunoaffinity column extraction and HPLC. AFM1 was detected in 125 (78%) of milk samples (ranging from < 1 ng/l to 23.5 ng/l; mean level: 6.28 ng/l), in 49 (53%) of dry milk samples (ranging from <1 ng/l to 79.6 ng/kg; mean level: 32.2 ng/kg) and in 73 (61%) of yoghurt samples (ranging from <1 ng/kg to 32.1 ng/kg; mean level: 9.06 ng/kg). Altogether, only four samples of dry milk were over the legal limits established by the EC in 1999. It is concluded that during 1996, despite the widespread occurrence of AFM1, mean contamination levels in dairy products sold in Italy were not a serious human health hazard.     

Survey of the occurrence of Aflatoxin M1 in ovine milk by HPLC and its confirmation by MS

During the period of October-July 2000, 240 samples of dairy ewes milk, obtained from farms of Enna (Sicily, Italy), were checked for Aflatoxin M(1) (AFM(1)) by HPLC using a fluorimetric detector. The limit of detection and the limit of quantification were 250 ng/L for AFM(1). All the positive milk samples for AFM(1) were confirmed by LC-MS. AFM(1) was detected in 81% of milk samples, ranging from 2 to 108 ng/L. Three samples were over the legal limits (50 ng/L). Mean contamination of samples obtained from stabulated ewes was higher than that from grazing ewes (35.27 vs. 12.47 ng/L). Furthermore, samples collected in the period September-October showed higher contamination than samples collected during the other months (42.68 vs. 10.55 ng/L). Both differences are related to the administration of compound feed. Based on current toxicological knowledge we concluded that the AFM(1) contamination levels recorded in ewe milk did not present a serious human health hazard. However, as ewe milk is exclusively used to produce cheese due to its higher protein content, and also considering the preferential binding of AFM(1) to casein during coagulation of milk, a potentially high concentration effect could occur, thus the surveillance of contamination levels should be more continuous and widespread.     

Aflatoxin M1 and ochratoxin A in raw bulk milk from French dairy herds

Mycotoxins in milk are a public health concern and have to be regularly monitored. A survey on the presence of aflatoxin M₁ (AFM₁) and ochratoxin A (OTA) in raw bulk milk was conducted in 2003 in the northwest of France, the main French milk-producing basin. Randomly selected farms (n = 132) were characterized by a diet based on corn silage and containing a large proportion of on-farm produced cereals, feeding sources that are frequently contaminated by mycotoxins. Farms were surveyed twice in winter and in summer. At each sampling time, a trained surveyor completed a questionnaire recording farm management procedures and production traits. The AFM₁ was found in 3 out of 264 samples but at levels (26 ng/L or less) that are below the European legislation limit of 50 ng/L. Traces of AFM₁ (less than 8 ng/L) were also found in 6 other samples. The OTA was detected in 3 samples also at low levels, 5 to 8 ng/L. Farms that tested positive to the presence of mycotoxins, 12 in total including 6 farms that had traces of AFM₁, differed from negative farms by a more extensive use of total mixed rations, 58 vs. 27%. In addition, the positive farms tended to have lower milk yields. Although the incidence of milk contamination with AFM₁ and OTA at the farm level was low during the period studied, production and management data from the surveyed farms suggest a link between feeding management practices and mycotoxin contamination.     

Aflatoxin M1 in yoghurts in Portugal

Aflatoxin M1 (AFM1) may occur in milk and milk products, resulting from the ingestion of aflatoxin B1 in feedstuffs by dairy cow. Ninety-six samples of commercial yoghurts (48 natural yoghurts and 48 yoghurt with pieces of strawberries) that are produced in Portugal were analyzed for the presence of AFM1 by immunoaffinity column extraction and high-performance liquid chromatography (HPLC). The limit of detection was 10 ng/kg. The recoveries of AFM1 from the samples spiked at levels of 10.0, 50.0, 100.0 and 150.0 ng/kg were 88.0%, 91.0%, 93.0% and 99.0%, respectively. AFM1 was detected in 18 (18.8%) of yogurt samples ranging from 19 to 98 ng/kg, and 78 samples (81.2%) did not reveal the presence of the toxin. Of the 48 natural yoghurts tested, only two (4.2%) were contaminated with 43 and 45 ng/kg of AFM1. Of the 48 yoghurts with pieces of strawberries tested, 16 samples (33.3%) contained levels ranging from 19 to 98 ng/kg; six samples (12.5%) were contaminated with low levels ranging from 19 to 35 ng/kg; four samples (8.3%) were contaminated with levels ranging from 36 to 50 ng/kg, two samples (4.2%) with levels of 51 and 65 ng/kg and four samples (8.3%) presented high contamination levels, from 90 to 98 ng/kg. This paper reports the data of the first survey on the presence of AFM1 in yoghurt in Portugal.     

Short communication: Aflatoxin M1 in dairy products sold in Şanlıurfa,Turkey

The aim of this study was to detect the presence of aflatoxin M₁ (AFM1) in samples of raw milk (n = 38), UHT milk (n = 12), white pickled cheese (n = 50), and yogurt (n = 50) collected from the ?anl?urfa city markets and locally produced dairy products by ELISA. The mean contamination rates were 56.74 ± 40.32, 43.1 ± 23.19, 103.2 ± 29.13, and 55.28 ± 12.68 ng/kg, respectively, for raw milk, UHT milk, white pickled cheese, and yogurt. According to the data, 21 (55%) raw milk, 3 (25%) UHT milk, 10 (20%) white pickled cheese, and 10 (20%) yogurt samples were contaminated with AFM1 over the acceptable levels (≥50 ng/kg), ranging from 0.82 to 130.89 ng/kg. None of the white pickled cheese samples contained AFM1 levels above the Turkish legal limit (250 ng/kg). Consequently, the AFM1 contamination levels determined in this study in white pickled cheese were not considered to pose a serious public health hazard. However, the AFM1 levels in raw and UHT milk and yogurt samples indicate an increased human health risk in Turkey related to high aflatoxin levels. Therefore, milk and dairy products have to be monitored by the Turkish public health authorities continuously to detect AFM1 contamination.     

Survey of Romanian slaughtered pigs for the occurrence of mycotoxins ochratoxins A and B, and zearalenone

Blood serum, kidney, liver and muscle sample per animal were collected from slaughtered pigs (n = 52). The samples were analysed for ochratoxin A (OTA) and B (OTB) by HPLC methods. Zearalenone (ZEA) in serum was analysed by enzyme immunoassay. A total of 98% serum samples were OTA positive in the range of 0.05-13.4 ng/ml and 85% contained under 5 ng OTA/ml. The incidences of OTA in kidney and liver were very similar (79%, 75%) with mean levels of 0.54 ng/g and 0.16 ng/g, respectively. The lowest incidence (17%) and the lowest mean level contamination (0.15 ng/g) were in muscle samples. The mean distribution in tissues followed the pattern serum > kidney > liver > muscle (100%: 26%: 8.5%: 2.57%). No kidney, liver or muscle sample was found OTA positive above the maximum admitted limit in Romania (5 ng/g). No sample was found to be positive for OTB. A very similar OTA contamination (mean = 4.19 ng/ml, coefficient of variation = 34.4%) was observed in the serum samples (n = 10) collected from the same farm. A possible difference in regional distribution of OTA in Romania is suggested. Zearalenone was detected only in 17.3% of the serum samples with a maximum concentration of 0.96 ng/ml. This study shows the presence of OTA and ZEA in Romanian slaughtered pigs at levels comparable to those reported in other countries.     

Presence of aflatoxin M1 in pasteurized milk from Morocco

Fifty four samples of pasteurized milk produced by five different dairies from Morocco were surveyed for the presence of aflatoxin M1 (AFM1) using immunoaffinity columns and liquid chromatography coupled to fluorescence detection. Confirmation of AFM1 identity in positive samples was based on the formation of AFM1 hemi-acetal derivative (AFM2a) after derivatization with trifluoracetic acid. Analytical results showed that 88.8% of the samples were contaminated with AFM1; 7.4% being above the maximum level of 0.05 microg/L set by the Moroccan and European regulations for AFM1 in liquid milk. The incidence of AFM1 in milk from these dairies was 100, 92.3, 90, 83.3 and 77.7% respectively, with AFM1 levels ranging from 0.001 to 0.117 microg/L and a mean value of 0.0186 microg/L. Based on the results presented in this study, the estimated daily intake of AFM1 was 3.26 ng/person/day. In this work, data on the natural occurrence of AFM1 in pasteurized milk produced in Morocco is presented for the first time.     

北京地区啤酒中赭曲霉毒素A污染调查及暴露评估

目的了解北京地区出售的啤酒中赭曲霉毒素A(ochratoxinA,OTA)的污染水平,评估本地居民对啤酒中OTA的暴露水平。方法采用免疫亲和柱-高效液相色谱-荧光检测器检测法(IAC-HPLC-FD)对啤酒样品中OTA污染水平进行测定,并参照欧盟膳食暴露评估方法进行评估。结果在检测的87份啤酒样品中,阳性样品39份,阳性率是44.83%。OTA的污染水平范围0.010～0.093ng/ml,平均污染水平为0.015ng/ml,阳性样品的平均污染水平为0.026ng/ml。居民每周从啤酒中摄入的OTA范围为0.01～0.50ng/kgBW。结论与其他国家啤酒中OTA污染水平相比,北京地区的污染水平较低。居民啤酒OTA暴露量远低于JECFA提出的100ng/kgBW的PTWI和欧盟提出的5ng/kgBW的TDI。     

Mycotoxin detection in infant formula milks in Italy

After birth, infant formulas constitute an important or often sole food source for infants during the first months of life. In this study, a survey on the presence of aflatoxin M₁ (AFM1) and ochratoxin A (OTA) in the 14 leading brands of infant formulas marketed in Italy was conducted. Mycotoxins were determined by immunoaffinity column clean-up and high-performance liquid chromatography (HPLC) with fluorescence detection. AFM1 was found in two of 185 samples, but at levels below the European legislation limit of 25 ng l^?1. OTA was detected in 133 (72%) samples (range = 35.1–689.5 ng l^?1). It has been observed that OTA contamination was 80% in the ready-to-use preparations and 63% in the powdered samples. The Scientific Committee for Food (SCF) reviewed the toxicology on OTA and concluded that it would be prudent to reduce exposure to OTA ensuring that exposure is towards the lower end of the range of tolerable daily intakes of 1.2–14 ng kg^?1 body weight day^?1. OTA was also evaluated by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) and a provisional tolerable weekly intake (PTWI) of 100 ng kg^?1 body weight was established. The OTA levels in pre-term ready-to-use infant formulas were sufficient to cause a higher OTA intake than the suggested TDI. The results point out the need to perform controls for prevention programmes especially when attempting to identify risk markers of the infant feed quality.     

Contribution to the study of ochratoxin A in Spanish wines.

With the aim of assessing ochratoxin A (OTA) contamination in wines from a Spanish northern region and the influence of harvest conditions, the following samples were analysed: 40 wines (28 red and 12 white) obtained from grapes cultivated in three different places of the northern Spanish region of Navarra, but in different years, 20 samples in 1997 and 20 in 1998. Wine samples were provided by a viticultural experimental station with very consistent and controlled cultural and enological practices. A high-performance liquid chromatography (HPLC) method with fluorescence detection and immunoaffinity columns was employed (LOD = 0.05 ng ml(-1); method recovery = 101%). Eighty five per cent of the samples from 1997 showed OTA levels >0.05 ng ml(-1) (range 0.056-0.316 ng ml(-1)) and 15% of the samples from 1998 showed OTA levels above the LOD (range 0.074-0.193 ng ml(-1)). Averages detected in 1997 positive samples were 0.185 ng OTA ml(-1) wine (SD = 0.023) in white wine samples (n = 6) and 0.160 ng ml(-1) (SD = 0.119) in the red wine samples (n = 11). These differences between contamination by OTA in the samples from the two different years were attributed to the different quality of the grapes, due to the bad climate in 1997. The possibility of the loss of the mycotoxin was excluded by the analysis of OTA in contaminated wine during 12 months. This study showed that OTA is stable in wine for at least 1 year.     

Aflatoxin M1 in milk in Hisar city,Haryana, India and risk assessment

ABSTRACT

Aflatoxin M1 (AFM1) contamination in 150 samples of milk, sold in market of Hisar city of Haryana, India, was investigated by using High Pressure Liquid Chromatography (HPLC). Out of these, 40 samples contained AFM1 at a concentration below the limit of detection (LOD) of 0.052 μg/kg. Among the AFM1 contaminated samples, 46 raw milk samples contained a concentration above the LOD but less than the limit of quantitation (LOQ), whereas 64 samples were above the LOQ. Of these samples, 31 exceeded the maximum limit of 0.5 μg/kg prescribed by FSSAI, India. Based on this study, the dietary intake of AFM1 for adults through consumption of milk was estimated. The results indicated that AFM1 contamination can be a food safety issue for raw and pasteurised milk consumed in India. Therefore, there is a need for a national monitoring programme to control the level of mycotoxins in milk.     

The transfer of aflatoxin M1 in milk of ewes fed diet naturally contaminated by aflatoxins and effect of inclusion of dried yeast culture in the diet

An experiment was carried out to investigate 1) the transfer of aflatoxin M1 (AFM1) into the milk of dairy ewes fed diets naturally contaminated with aflatoxin B1 (AFB1); 2) the effect of the addition of dried yeast culture in the diet on this transfer; and 3) the alteration of enzymatic activities in the liver of ewes fed diets contaminated with AFB1. Twenty-four Sarda dairy ewes were divided in 4 groups and fed a concentrate mix containing 4 amounts of wheat meal naturally contaminated with aflatoxins. The diet of the control group had no wheat meal, whereas that of treated groups had low, medium, or high amounts of contaminated wheat, which corresponded to 1.13, 2.30, and 5.03 μg of AFB1/kg of feed, respectively. The experiment lasted 14 d. On d 8 to 14 from the beginning of the trial, 12 g/d of a commercial dried yeast product (DYP) of Kluyveromyces lactis was added to the diet of each ewe. The AFM1 concentration in individual milk samples and the blood serum metabolites were measured periodically. The presence of AFM1 was first detected in milk on d 1 of administration, and then its concentration increased and approached a steady-state condition on d 3 simultaneously in all treated groups. The AFM1 in milk at the steady-state condition, which was linearly related to the AFB1 intake, was 39.72, 50.38, and 79.29 ng/L in the low-aflatoxin, medium-aflatoxin, and high-aflatoxin groups, respectively. The AFM1 concentration in milk of the high-aflatoxin group was approximately 1.5-fold greater than the European Commission maximum tolerance level (50 ng/kg). The addition of DYP to the diet did not affect the AFM1 concentration in milk. After the withdrawal of the contaminated concentrate mix, the AFM1 mean concentrations decreased quickly and were no longer detected after 3 d in all treated groups. Daily milk yield and composition did not differ because of aflatoxin treatment. Blood serum parameters (creatinine, glutamic oxalacetic transaminase, glutamic pyruvic transaminase, gamma glutamyl transferase, alkaline phosphatase, lactate dehydrogenase, cholesterol, protein, urea, calcium, and phosphorus) were not influenced by AFB1 intake. Therefore, the effect of DYP on certain blood parameters (gamma glutamyl transferase, urea, creatinine, and calcium) could not be attributed to amelioration of the aflatoxin-contaminated diet. In conclusion, diet contamination by AFB1 near the European Union tolerance level (0.005 mg/kg) in complete feed for dairy animals (e.g., high-aflatoxin group) can result in an AFM1 milk concentration higher than the European Commission maximum tolerance level. Transfer of aflatoxin from feed to milk was not affected by dietary addition of a commercial DYP.     

Ochratoxin A in rice on the Moroccan retail market

One hundred (100) samples of rice purchased from retail markets in five different cities (Rabat, Témara, Salé, Casablanca and Méknès) in Morocco from January to October 2006 were surveyed for the presence of ochratoxin A (OTA) using Accelerated Solvent Extraction (ASE) coupled to liquid chromatography with fluorescence detection. The identification of OTA in positive rice samples was confirmed by methyl ester derivatization. Analytical results showed a frequency of contamination of 26% of total analyzed rice samples. The percentage of contamination of samples was 24, 26.6, 16.6, 27.7 and 30% in Rabat, Témara, Méknès, Salé and Casablanca respectively. Levels of OTA in positive samples ranged between 0.08 and 47 ng/g. The average contamination of all analyzed samples was 3.5 ng/g. The highest frequency of positive samples (30%) and the most contaminated sample (47 ng/g) was found in a sample from Casablanca city. 14 out of 100 total samples exceeded the maximum level of 5 ng/g set by European regulations for OTA in cereals. Based in the results presented in this study, the estimated daily intake of OTA in rice was 0.32 ng/kg bw/day for Moroccan consumers.     

Occurrence of aflatoxin M1 in retail milk samples from Bogotá, Colombia.

A study was conducted to establish the occurrence and levels of contamination of aflatoxin M1 (AFM1) in retail milk from Bogotá, Colombia. A total of 241 samples were analysed during 2004 and 2005. Samples were cleaned up by an immunoaffinity column and AFM1 was quantified by liquid chromatography with fluorescence detection. A total of 69.2 and 79.4% of the samples analysed during 2004 and 2005, respectively, were found to contain levels of AFM1 above 10 ng l(-1). Levels of contamination ranged from 10.7 to 213.0 ng l(-1) in 2004, and from 10.6 to 288.9 ng l(-1) in 2005. Despite the high incidence of AFM1 found in the milk samples analysed, all samples complied with current local regulations, which allow AFM1 content in milk up to 400 ng l(-1). However, due to the high incidence of AFM1 in milk found in the present study, it is recommended that a permanent surveillance programme be established for milk consumed in Bogotá in order to prevent milk lots containing levels above the regulatory level entering the food chain.     

Survey of aflatoxin M1 in cow's milk for human consumption in Kerman Province of Iran

The aim of this study was to assess levels of aflatoxin M₁ (AFM1) in milk samples from Kerman, Iran. AFM1 was detected in 72 samples, ranging in concentration from <0.01 to 0.41?µg?l^?1. The samples were analyzed using immunoaffinity column for clean-up and HPLC for determining AFM1. Milk samples were collected from six dairy farms. AFM1 was found in ～50% of the milk samples. The average level of AFM1 was below the tolerance limit (0.05?µg?l^?1), but 50% of the samples had greater levels than the maximum tolerance limit accepted by EU and the Iranian national standard. The method detection limit and limit of quantification were 0.01 and 0.03?µg?l^?1, respectively, and recovery of the method was 87%. The results showed that AFM1 contamination is a serious problem for public health. To achieve a low level of AFM1 in milk, cattle feed must be monitored regularly for aflatoxin contamination and protected from fungal contamination as much as possible.     

Study of ochratoxin A content in South Moravian and foreign wines by the UPLC method with fluorescence detection

In 2009–2010, 72 wine samples of Moravian and foreign origin were analysed for ochratoxin A contamination. A fast analytical method based on immunoaffinity column clean-up and followed by the ultra performance liquid chromatography coupled to fluorescence detection was used. LOD and LOQ values were 0.3 and 1.0 ng/L. Ochratoxin A was detected in 11% of Moravian wines and the detected OTA level ranged from 1.2 to 71.2 ng/L. In foreign wines, OTA level ranged from 1.6 to 227.0 ng/L. The values of OTA in all studied samples were significantly below the maximum allowable limit, 2.0 μg/kg, set by the European Union.     

Presence of ochratoxin A in human milk in relation to dietary intake.

Individual and geographical variations in ochratoxin A (OA) levels in human blood and milk samples may be due to differences in dietary habits. The purpose of this study was to examine the relationship between OA contamination of human milk and dietary intake. Human milk samples were collected from 80 Norwegian women. The usual food intake during the last year was recorded using a quantitative food frequency questionnaire. The concentration of OA in the human milk was determined by HPLC (detection limit 10 ng/l). Seventeen (21%) out of 80 human milk samples contained OA in the range 10-182 ng/l. The women with a high dietary intake of liver paste (liverwurst, liver paté) and cakes (cookies, fruitcakes, chocolate cakes, etc.) were more likely to have OA-contaminated milk. The risk of OA contamination was also increased by the intake of juice (all kinds). In addition, the results indicate that breakfast cereals, processed meat products, and cheese could be important contributors to dietary OA intake. OA contamination of the milk was unrelated to smoking, age, parity, and anthropometric data other than body weight.     

Excretion pattern of aflatoxin M1 in milk of goats fed a single dose of aflatoxin B1

The feedstuffs used in dairy animals must be able to give consumers confidence about the wholesomeness of milk with regard to aflatoxin contamination. The aim of this study was to determine the excretion patterns of aflatoxin M(1) (AFM1) in the milk of dairy goats fed a single dose of pure aflatoxin B(1) (AFB1), which can occasionally occur if feeds are infected by hot-spot growth of molds that produce aflatoxins. Five dairy goats in midlactation were administered 0.8 mg of AFB1 orally. Individual milk samples were collected for 84 h after AFB1 dosage. Aflatoxin M(1) was found in milk in the highest concentration. In all goats, AFM1 was not detected in milk before AFB1 administration, but was detected in the first milking following AFB1 administration. The excretion pattern of AFM1 concentration in milk was very similar in all goats even if the values of the concentration differed between animals. The peak values for AFM1 concentration in milk was observed in milk collected during the milking at 3 and 6h. After the peak, the AFM1 in milk disappeared with a trend that fitted well a monoexponential decreasing function, and the toxin was not detected after 84 h. Only about 0.17% of the amount of AFB1 administered was detected as AFM1 in milk, and about 50% of this was excreted in the first liter of milk yielded after AFB1 intake. Correct procedures to prevent growth of molds, and consequent AFB1 contamination, on the feedstuffs for lactating goats represent the key to providing consumers a guarantee that milk is not contaminated by AFM1.     

Determination of ochratoxin A in wine by immunoaffinity cleanup and liquid chromatography tandem mass spectrometry

A simple and accurate method has been developed for determining ochratoxin A (OTA), using an immunoaffinity column for cleanup and liquid chromatography-tandem mass spectrometry for identification and quantification. Wine samples were diluted with a solution containing polyethylene glycol 8000 and sodium hydrogen carbonate, filtered through a glass microfiber filter, and cleaned up on an immunoaffinity column. OTA was then eluted with methanol-acetic acid (98:2) and analyzed by liquid chromatography-tandem mass spectrometry. The average recoveries of OTA from red and white wines were 95 and 96.7% (spiked OTA level was 0.05 ng/ml) and repeatabilities (relative standard deviation) were 3.8 and 2.4%, respectively. The detection limit was 0.0003 ng/ml based on the signal-to-noise ratio in wine of 3:1. Analysis of 74 samples of domestic and imported wines showed OTA levels ranging from < 0.0003 to 0.82 ng/ml, with an incidence of contamination of 92.1% for red wines, and < 0.0003 to 0.51 ng/ml, with an incidence of contamination of 77.8% for white wines. These detection rates were higher than those rates of past reports of OTA contamination in wine, due to the high sensitivity of this method. However, all samples analyzed in this study complied with European Union regulations. It is concluded that this method is a useful tool for the quality assurance of wine.     

Impact of industrial treatments on ochratoxin A content in artificially contaminated cocoa beans

Ochratoxin A (OTA) is a mycotoxin mainly produced by mould species of the genera Aspergillus and Penicillium, which grow on a variety of agricultural products. OTA-contaminated foodstuffs pose a major health hazard to consumers, including human and animal. In Côte d’Ivoire, numerous studies are being carried out to find the best way of preventing OTA contamination of cocoa raw material. The objectives of this investigation were to assess the impact of industrial treatment on OTA content in cocoa-derived products. Samples of cocoa pods were prepared under specific conditions promoting fungal proliferation on cocoa beans before processing. The beans underwent the usual industrial treatments – roasting, shelling, crushing, pressing and additive addition – and samples were taken at each stage. OTA was extracted with a methanol/3% sodium hydrogen carbonate solution and purified using an immunoaffinity column prior to HPLC analysis with fluorescence detection. OTA was detected in artificially contaminated cocoa beans at levels ranging from 3.4 to 44.7 µg kg^?1 with a mean value of 22.9 ± 3.6 µg kg^?1. OTA was mainly concentrated in the shell (93%). Roasting, shelling and additive addition significantly decreased levels of OTA by 24–40, 76 and 52%, respectively, with an overall reduction of ～91%. These results indicate that industrial processing of cocoa has a real impact on the reduction of OTA in final cocoa products.